Original Article

Effects of Continuous and Interrupted Forces on Gene Transcription in Periodontal Ligament Cells in Vitro

Abstract

The biological mechanisms of tooth movement are based on the response of periodontal tissues to mechanical forces. The final result of these responses is remodeling of the extracellular matrix. Tissue reactions may vary depending upon the type, magnitude and duration of the applied forces. The purpose of the present study was to analyze the effects of centrifugal force on the transcription of collagen type-I (Col-I), matrix metalloproteinase-1 (MMP-1), and tissue inhibitor of metalloproteinase- 1 (TIMP-1) genes in human periodontal ligament (PDL) fibroblasts. Human fibroblasts obtained from the PDL were cultured and subjected to centrifugal forces (36.3 g/cm2) for 30, 60 and 90 min continuously. This was also carried out interruptedly, three times for 30 min and six times for 15 min. The mRNAs encoding for Col-I, MMP-1, and TIMP-1 were quantified using RT-PCR. The mRNA levels of Col-I and MMP-1 were increased when continuous force was applied for 30 min and 60 min respectively. The interrupted force had almost no effect on Col-I, MMP-1 and TIMP-1 genes. These results indicate that continuous forces may have a greater effect in inducing gene expression during the remodeling process of PDL compared to interrupted forces with short rest periods.

Meikle M. The tissue, cellular, and molecular regulation of orthodontic tooth movement: 100 years after Carl Sandstedt. Eur J Orthod 2006;28(3):221-40.

Nahm DS, Kim HJ, Mah J, Baek SH. In vitro expression of matrix metalloproteinase-1, tissue inhibitor of metalloproteinase-1 and transforming growth factor-beta1 in human periodontal ligament fibroblasts. Eur J Orthod 2004;26(2):129-35.

Cawston TE, Wilson AJ. Understanding the role of tissue degrading enzymes and their inhibitors in development and disease. Best Pract Res Clin Rheumatol 2006;20(5):983-1002.

Nagase H, Visse R, Murphy G. Structure and function of matrix metalloproteinases and TIMPs. Cardiovasc Res 2006;69(3):562-73.

Verstappen J, Von den Hoff JW. Tissue inhibitors ofmetalloproteinases (TIMPs): their biological functions and involvement in oral disease. J Dent Res 2006;85(12):1074-84.

Blavier L, Lazaryev A, Groffen J, Heisterkamp N, DeClerck YA, Kaartinen V. TGF-β3-induced= palatogenesis requires matrix metalloproteinases. Mol Biol Cell 2001;12(5):1457-66.

Brown TD. Techniques for mechanical stimulation of cells in vitro: a review. J Biomech 2000;33(1):3-14.

Carano A, Siciliani G. Effects of continuous and intermittent forces on human fibroblasts in vitro. Eur J Orthod 1996;18(1):19-26.

Bolcato-Bellemin AL, Elkaim R, Abehsera A, Fausser JL,Haikel Y, Tenenbaum H. Expression of mRNAs encoding for alpha and beta integrin subunits, MMPs, and TIMPs in stretched human periodontal ligament and gingival fibroblasts. J Dent Res 2000;79(9):1712-6.

Redlich M, Roos H, Reichenberg E, Zaks B, Grosskop A, Bar Kana I, Pitaru S, Palmon A. The effect of centrifugal force on mRNA levels of collagenase, collagen type-I, tissue inhibitors of metalloproteinases and beta-actin in cultured human periodontal ligament fibroblasts. J Periodontal Res 2004;39(1):27-32.

Long P, Liu F, Piesco NP, Kapur R, Agarwal S. Signaling by mechanical strain involves transcriptional regulation of proinflammatory genes in human periodontal ligament cells in vitro. Bone 2002;30(4):547-52.

Somerman MJ, Archer SY, Imm GR, Foster RA. Acomparative study of human periodontal ligament cells and gingival fibroblasts in vitro. J Dent Res 1988;67(1):66-70.

Davidovitch Z. Tooth movement. Crit Rev Oral Biol Med 1991;2(4):411-50.

Schwaller J, Pabst T, Bickel M, Borisch B, Fey MF, Tobler A. Comparative detection and quantitation of human CDK inhibitor mRNA expression of p15INK4B, p16INK4A, p16beta, p18INK4C, p19INK4D, p21WAF1, p27KIP1 and p57KIP2 by RT-PCR using a polycompetitive internal standard. Br J Haematol 1997;99(4):896-900.

Basdra EK, Komposch G. Osteoblast-like properties ofhuman periodontal ligament cells: an in vitro analysis. Eur J Orthod 1997;19(6):615-21.

Myokai F, Oyama M, Nishimura F, Ohira T, Yamamoto T, Arai H, Takashiba S, Murayama Y. Unique genes inducedby mechanical stress in periodontal ligament cells. J Periodontal Res 2003;38(3):255-61.

Redlich M, Reichenberg E, Harari D, Zaks B, Shoshan S,Palmon A. The effect of mechanical force on mRNA levels of collagenase, collagen type I, and tissue inhibitors of metalloproteinases in gingivae of dogs. J Dent Res 2001;80(12):2080-4.

Apajalahti S, Sorsa T, Railavo S, Ingman T. The in vivo levels of matrix metalloproteinase-1 and -8 in gingival crevicular fluid during initial orthodontic tooth movement. J Dent Res 2003;82(12):1018-22.

Takahashi I, Nishimura M, Onodera K, Bae JW, Mitani H, Okazaki M, Sasano Y, Mitani H. Expression of MMP-8 and MMP-13 genes in the periodontal ligament during tooth movement in rats. J Dent Res 2003;82(8):646-51.

Ingman T, Apajalahti S, Mäntylä P, Savolainen P, Sorsa T. Matrix metalloproteinase-1 and -8 in gingival crevicular fluid during orthodontic tooth movement: a pilot study during 1 month of follow-up after fixed appliance activation. Eur J Orthod 2005;27(2):202-7.

Cantarella G, Cantarella R, Caltabiano M, Risuglia N, Bernardini R, Leonardi R. Levels of matrix metalloproteinases 1 and 2 in human gingival crevicular fluid during initial tooth movement. Am J Orthod Dentofacial Orthop 2006;130(5):568.e11-6.

Wescott DC, Pinkerton MN, Gaffey BJ, Beggs KT, MilneTJ, Meikle MC. Osteogenic gene expression by human periodontal ligament cells under cyclic tension. J Dent Res 2007;86(12):1212-6.

Nakao K, Goto T, Gunjigake KK, Konoo T, Kobayashi S, Yamaguchi K. Intermittent force induces high RANKL expression in human periodontal ligament cells. J Dent Res 2007;86(7):623-8.

Alaqeel SM, Hinton RJ, Opperman LA. Cellular response to force application at craniofacial sutures. Orthod Craniofac Res 2006;9(3):111-22.

Lee KJ, Park YC, Yu HS, Choi SH, Yoo YJ. Effects of continuous and interrupted orthodontic force on interleukin-1beta and prostaglandin E2 production in gingival crevicular fluid. Am J Orthod Dentofacial Orthop 2004;125(2):168-77.

Gilsanz V, AL Wren T, Sanchez M, Dorey F, Judex S, Rubin C. Low-level, high-frequency mechanical signals enhance musculoskeletal development of young women with low BMD. J Bone Miner Res 2006;21(9):1464-74.

Howard PS, Kucich U, Taliwal R, Korostoff JM. Mechanical forces alter extracellular matrix synthesis by human periodontal ligament fibroblasts. J Periodontal Res 1998;33(8):500-8.

Rubin CT, Lanyon LE. Regulation of bone formation by applied dynamic loads. J Bone Joint Surg Am 1984;66(3):397-402.

Robling AG, Burr DB, Turner CH. Recovery periods restore mechanosensitivity to dynamically loaded bone. J Exp Biol 2001;204(Pt 19):3389-99.

Chiba M, Mitani H. Cytoskeletal changes and system of regulation of alkaline phosphatase activity in human periodontal ligament cells induced by mechanical stress. Cell Biochem Funct 2004;22(4):249-56

Files
IssueVol 49, No 10 (2011) QRcode
SectionOriginal Article(s)
Keywords
Gene expression Stress mechanical Periodontal ligament Reverse transcriptase polymerase chain reaction

Rights and permissions
Creative Commons License This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
How to Cite
1.
Hacopian N, Hosseinzadeh Nik T, Ghahremani MH, Rahimi HR, Ostad SN. Effects of Continuous and Interrupted Forces on Gene Transcription in Periodontal Ligament Cells in Vitro. Acta Med Iran. 1;49(10):643-649.